A new Real-Time-RT-PCR for quantitation of human endogenous retroviruses type K (HERV-K) RNA load in plasma samples: increased HERV-K RNA titers in HIV-1 patients with HAART non-suppressive regimens

J Virol Methods. 2006 Sep;136(1-2):51-7. doi: 10.1016/j.jviromet.2006.03.029. Epub 2006 May 6.

Abstract

Viral components of the human endogenous retroviruses type K (HERV-K) have been largely detected in plasma from HIV-1 infected individuals. A Sybr Green Real-Time RT-PCR approach was optimized for detection and quantitation of HERV-K RNA titers in plasma samples using the iCycler technology. The method detected 1000 HERV-K RNA copies/mL of plasma sample. The Intra- and Inter-assay performance revealed a coefficient of variations that ranged from 0.2 to 2.46%, demonstrating accuracy and reproducibility. We quantified the HERV-K RNA load in 20 HIV-1 patients receiving highly active antiretroviral therapy (HAART). We found increased HERV-K RNA titers in patients with non-suppressive HAART (patients who may develop drug-resistance and/or received suboptimal therapeutic doses), compared to suppressive regimens (p < 0.001). HERV-K RNA was not detected in HCV-1 positive or seronegative controls. Sequencing of Real-Time RT-PCR products revealed particular HERV-K subtypes activated in the HIV-1 infection. The application of this assay could expand the understanding of the role of HERV-K in the HIV-1 infection and others pathological conditions.

Publication types

  • Evaluation Study
  • Research Support, N.I.H., Extramural

MeSH terms

  • Analysis of Variance
  • Antiretroviral Therapy, Highly Active*
  • Base Sequence
  • Cloning, Molecular
  • Endogenous Retroviruses / genetics
  • Endogenous Retroviruses / isolation & purification*
  • Fluorescence
  • HIV Infections / complications*
  • HIV Infections / drug therapy*
  • HIV Infections / virology
  • HIV-1*
  • Humans
  • Molecular Sequence Data
  • Organic Chemicals
  • RNA, Viral / blood
  • Reproducibility of Results
  • Reverse Transcriptase Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity
  • Staining and Labeling
  • Viral Load*

Substances

  • Organic Chemicals
  • RNA, Viral
  • SYBR Green I